Tenofovir alafenamide hemifumarate

ABSTRACT

A hemifumarate form of 9-[(R)-2-[[(S)-[[(S)-1-(isopropoxycarbonyl)ethyl]amino]phenoxyphosphinyl]methoxy]propyl]adenine (tenofovir alafenamide), and antiviral therapy using tenofovir alafenamide hemifumarate (e.g., anti-HIV and anti-HBV therapies).

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of application Ser. No. 13/586,358filed Aug. 15, 2012, which in turn claims the benefit of priority fromU.S. Provisional Patent Application No. 61/524,224, filed Aug. 16, 2011,the content of each of which is hereby incorporated by reference hereinin its entirety.

BACKGROUND OF THE INVENTION Description of Related Art

U.S. Pat. Nos. 7,390,791 and 7,803,788 (the content of each of which isincorporated by reference herein in its entirety) describe certainprodrugs of phosphonate nucleotide analogs that are useful in therapy.One such prodrug is9-[(R)-2-[[(S)-[[(S)-1-(isopropoxycarbonyl)ethyl]amino]phenoxyphosphinyl]methoxy]propyl]adenine.This compound is also known by the Chemical Abstract name L-alanine,N—[(S)-[[(1R)-2-(6-amino-9H-purin-9-yl)-1-methylethoxy]methyl]phenoxyphosphinyl]-,1-methylethyl ester. U.S. Pat. Nos. 7,390,791 and 7,803,788 alsodisclose a monofumarate form of this compound and its preparation method(see, e.g., Example 4).

SUMMARY OF THE INVENTION

Described is a hemifumarate form of9-[(R)-2-[[(S)-[[(S)-1-(isopropoxycarbonyl)ethyl]amino]phenoxyphosphinyl]methoxy]propyl]adenine.The name for9-[(R)-2-[[(S)-[[(S)-1-(isopropoxycarbonyl)ethyl]amino]phenoxyphosphinyl]methoxy]propyl]adenineis tenofovir alafenamide. The hemifumarate form of tenofovir alafenamideis also referred to herein as tenofovir alafenamide hemifumarate.

In one embodiment of the invention is provided tenofovir alafenamidehemifumarate.

In another embodiment is provided tenofovir alafenamide hemifumarate,wherein the ratio of fumaric acid to tenofovir alafenamide is 0.5±0.1,or 0.5±0.05, or 0.5±0.01, or about 0.5.

In one embodiment is provided tenofovir alafenamide hemifumarate in asolid form.

In one embodiment is provided tenofovir alafenamide hemifumarate thathas an X-ray powder diffraction (XRPD) pattern having 2theta values of6.9±0.2° and 8.6±0.2°. In another embodiment is provided tenofoviralafenamide hemifumarate wherein the XRPD pattern comprises 2thetavalues of 6.9±0.2°, 8.6±0.2°, 11.0±0.2°, 15.9±0.2°, and 20.2±0.2°.

In one embodiment is provided tenofovir alafenamide hemifumarate thathas a differential scanning calorimetry (DSC) onset endotherm of 131±2°C., or 131±1° C.

In one embodiment is provided a pharmaceutical composition comprisingtenofovir alafenamide hemifumarate and a pharmaceutically acceptableexcipient. In another embodiment is provided the pharmaceuticalcomposition, further comprising an additional therapeutic agent. In afurther embodiment, the additional therapeutic agent is selected fromthe group consisting of human immunodeficiency virus (HIV) proteaseinhibiting compounds, HIV nonnucleoside inhibitors of reversetranscriptase, HIV nucleoside inhibitors of reverse transcriptase, HIVnucleotide inhibitors of reverse transcriptase, HIV integraseinhibitors, and CCR5 inhibitors.

In one embodiment is provided a method for treating a humanimmunodeficiency virus (HIV) infection comprising administering to asubject in need thereof a therapeutically effective amount of tenofoviralafenamide hemifumarate. In another embodiment is provided a method fortreating an HIV infection comprising administering to a subject in needthereof a therapeutically effective amount of a pharmaceuticalcomposition comprising tenofovir alafenamide hemifumarate. In a furtherembodiment, the method comprises administering to the subject one ormore additional therapeutic agents selected from the group consisting ofHIV protease inhibiting compounds, HIV nonnucleoside inhibitors ofreverse transcriptase, HIV nucleoside inhibitors of reversetranscriptase, HIV nucleotide inhibitors of reverse transcriptase, HIVintegrase inhibitors, and CCR5 inhibitors.

In one embodiment is provided a method for treating a hepatitis B virus(HBV) infection comprising administering to a subject in need thereof atherapeutically effective amount of tenofovir alafenamide hemifumarate.In another embodiment is provided a method for treating an HBV infectioncomprising administering to a subject in need thereof a therapeuticallyeffective amount of the pharmaceutical composition comprising tenofoviralafenamide hemifumarate.

In one embodiment is provided a method for preparing a pharmaceuticalcomposition comprising combining tenofovir alafenamide hemifumarate anda pharmaceutically acceptable excipient to provide the pharmaceuticalcomposition.

In one embodiment is provided a method for preparing tenofoviralafenamide hemifumarate comprising subjecting a solution comprising asuitable solvent; fumaric acid; tenofovir alafenamide; and, optionally,one or more seeds of tenofovir alafenamide hemifumarate to conditionsthat provide for the crystallization of the fumaric acid and thetenofovir alafenamide. In one embodiment, the solvent comprisesacetonitrile. In another embodiment, the solution is subjected to atemperature in the range of from about 0° C. to about 75° C.

In one embodiment is provided tenofovir alafenamide hemifumarate for usein medical therapy.

In one embodiment is provided the use of tenofovir alafenamidehemifumarate for the prophylactic or therapeutic treatment of an HIVinfection. In another embodiment is provided the use of tenofoviralafenamide hemifumarate to treat an HIV infection. In a furtherembodiment is provided the use of tenofovir alafenamide hemifumarate forthe preparation or manufacture of a medicament for the treatment of anHIV infection. In another further embodiment is provided tenofoviralafenamide hemifumarate for use in treating an HIV infection.

In one embodiment is provided the use of tenofovir alafenamidehemifumarate for the prophylactic or therapeutic treatment of an HBVinfection. In another embodiment is provided the use of tenofoviralafenamide hemifumarate to treat an HBV infection. In a furtherembodiment is provided the use of tenofovir alafenamide hemifumarate forthe preparation or manufacture of a medicament for the treatment of anHBV infection. In another further embodiment is provided tenofoviralafenamide hemifumarate for use in treating an HBV infection.

In some embodiments of the invention, the methods of treating and thelike comprise administration of multiple daily doses. In otherembodiments, the methods of treating and the like compriseadministration of a single daily dose.

In one embodiment of the invention is provided a composition consistingessentially of tenofovir alafenamide hemifumarate.

BRIEF DESCRIPTIONS OF THE DRAWINGS

FIG. 1 shows the X-ray powder diffraction (XRPD) pattern of tenofoviralafenamide hemifumarate.

FIG. 2 shows a graph of the DSC analysis of tenofovir alafenamidehemifumarate.

FIG. 3 shows a graph of the thermogravimetric analysis (TGA) data fortenofovir alafenamide hemifumarate.

FIG. 4 shows a graph of the dynamic vapor sorption (DVS) analysis oftenofovir alafenamide hemifumarate.

DETAILED DESCRIPTION OF THE INVENTION

Specific values listed within the present description for radicals,substituents, and ranges are for illustration only; they do not excludeother defined values or other values within defined ranges for theradicals and substituents.

In one embodiment, there is provided a hemifumarate form of tenofoviralafenamide (i.e., tenofovir alafenamide hemifumarate). This form mayhave a ratio (i.e., a stoichiometric ratio or mole ratio) of fumaricacid to tenofovir alafenamide of 0.5±0.1, 0.5±0.05, 0.5±0.01, or about0.5, or the like.

In one embodiment, tenofovir alafenamide hemifumarate consists offumaric acid and tenofovir alafenamide in a ratio of 0.5±0.1.

In one embodiment, tenofovir alafenamide hemifumarate consistsessentially of fumaric acid and tenofovir alafenamide in a ratio of0.5±0.1.

In one embodiment, tenofovir alafenamide hemifumarate has an XRPDpattern comprising 2theta values of 6.9±0.2°, 8.6±0.2°, 10.0±0.2°,11.0±0.2°, 12.2±0.2°, 15.9±0.2°, 16.3±0.2°, 20.2±0.2°, and 20.8±0.2°.

In one embodiment, tenofovir alafenamide hemifumarate has an XRPDpattern comprising at least four 2theta values selected from 6.9±0.2°,8.6±0.2°, 10.0±0.2°, 11.0±0.2°, 12.2±0.2°, 15.9±0.2°, 16.3±0.2°,20.2±0.2°, and 20.8±0.2°.

In one embodiment, tenofovir alafenamide hemifumarate has a DSC onsetendotherm of 131±2° C., or 131±1° C.

In one embodiment, a tenofovir alafenamide hemifumarate compositioncomprises less than about 5% by weight of tenofovir alafenamidemonofumarate.

In one embodiment, a tenofovir alafenamide hemifumarate compositioncomprises less than about 1% by weight of tenofovir alafenamidemonofumarate.

In one embodiment, a tenofovir alafenamide hemifumarate compositioncomprises less than about 0.5% by weight of tenofovir alafenamidemonofumarate.

In one embodiment, a tenofovir alafenamide hemifumarate compositioncomprises no detectable tenofovir alafenamide monofumarate.

Tenofovir alafenamide (i.e., the compound9-[(R)-2-[[(S)-[[(S)-1-(isopropoxycarbonyl)ethyl]amino]phenoxyphosphinyl]methoxy]propyl]adenine)can be prepared as described in U.S. Pat. No. 7,390,791.

Selective Crystallization

In one embodiment, tenofovir alafenamide hemifumarate can be preparedusing selective crystallization. An example of a scheme for thispreparation method is as follows.

The method can be carried out by subjecting a solution comprising: a) asuitable solvent; b) fumaric acid; c) tenofovir alafenamide; and,optionally, d) one or more seeds comprising tenofovir alafenamidehemifumarate, to conditions that provide for the crystallization offumaric acid and tenofovir alafenamide. The starting solution cancontain the single diastereomer of tenofovir alafenamide or a mixture oftenofovir alafenamide and one or more of its other diastereomers (e.g.,GS-7339, as described in U.S. Pat. No. 7,390,791).

The selective crystallization can be carried out in any suitablesolvent. For example, it can be carried out in a protic solvent or in anaprotic organic solvent, or in a mixture thereof. In one embodiment, thesolvent comprises a protic solvent (e.g., water or isopropyl alcohol).In another embodiment, the solvent comprises an aprotic organic solvent(e.g., acetone, acetonitrile (ACN), toluene, ethyl acetate, isopropylacetate, heptane, tetrahydrofuran (THF), 2-methyl THF, methyl ethylketone, or methyl isobutyl ketone, or a mixture thereof). In oneembodiment, the solvent comprises ACN or a mixture of ACN and up toabout 50% methylene chloride (by volume). The selective crystallizationalso can be carried out at any suitable temperature, for example, atemperature in the range of from about 0° C. to about 70° C. In onespecific embodiment, the resolution is carried out at a temperature ofabout 0° C.

One major advantage of the hemifumarate form of tenofovir alafenamideover the monofumarate form is its exceptional capability to purgeGS-7339 (i.e.,9-[(R)-2-[[(R)-[[(S)-1-(isopropoxycarbonyl)ethyl]amino]phenoxyphosphinyl]methoxy]propyl]adenine;described in, e.g., U.S. Pat. No. 7,390,791), which is the majordiastereomeric impurity in the active pharmaceutical ingredient. Thus,the hemifumarate form of tenofovir alafenamide can be more readily andeasily separated from impurities than the monofumarate form. Other majoradvantages of tenofovir alafenamide hemifumarate over the monofumarateform include improved thermodynamic and chemical stability (includinglong-term storage stability), superior process reproducibility, superiordrug product content uniformity, and a higher melting point.

Tenofovir alafenamide hemifumarate is useful in the treatment and/orprophylaxis of one or more viral infections in man or animals, includinginfections caused by DNA viruses. RNA viruses, herpesviruses (e.g., CMV,HSV 1, HSV 2, VZV), retroviruses, hepadnaviruses (e.g., HBV),papillomavirus, hantavirus, adenoviruses and HIV. U.S. Pat. No.6,043,230 (incorporated by reference herein in its entirety) and otherpublications describe the antiviral specificity of nucleotide analogs,such as tenofovir disoproxil. Like tenofovir disoproxil, tenofoviralafenamide is another prodrug form of tenofovir, and can be used in thetreatment and/or prophylaxis of the same conditions.

Tenofovir alafenamide hemifumarate can be administered by any routeappropriate to the condition to be treated. Suitable routes includeoral, rectal, nasal, topical (including ocular, buccal, and sublingual),vaginal, and parenteral (including subcutaneous, intramuscular,intravenous, intradermal, intrathecal, and epidural). Generally,tenofovir alafenamide hemifumarate is administered orally, but it can beadministered by any of the other routes noted herein.

Accordingly, pharmaceutical compositions include those suitable fortopical or systemic administration, including oral, rectal, nasal,buccal, sublingual, vaginal, or parenteral (including subcutaneous,intramuscular, intravenous, intradermal, intrathecal, and epidural)administration. The formulations are in unit dosage form and areprepared by any of the methods well known in the art of pharmacy.

For oral therapeutic administration, the tenofovir alafenamidehemifumarate may be combined with one or more excipients and used in theform of ingestible tablets, buccal tablets, troches, capsules, elixirs,suspensions, syrups, wafers, and the like. Such pharmaceuticalcompositions and preparations will typically contain at least 0.1% oftenofovir alafenamide hemifumarate. The percentage of this activecompound in the compositions and preparations may, of course, be variedand may conveniently be between about 2% to about 60% or more of theweight of a given unit dosage form. The amount of active compound insuch therapeutically useful pharmaceutical compositions is preferablysuch that an effective dosage level will be obtained upon administrationof a single-unit dosage (e.g., tablet). Other dosage formulations mayprovide therapeutically effective amounts of tenofovir alafenamidehemifumarate upon repeated administration of subclinically effectiveamounts of the same. Preferred unit dosage formulations include thosecontaining a daily dose (e.g., a single daily dose), as well as thosecontaining a unit daily subclinical dose, or an appropriate fractionthereof (e.g., multiple daily doses), of tenofovir alafenamidehemifumarate.

Pharmaceutical compositions suitable for oral administration may bepresented as discrete units such as capsules, cachets, or tablets, eachcontaining a predetermined amount of tenofovir alafenamide hemifumarate;as a powder or granules; as a solution or a suspension in an aqueousliquid or a nonaqueous liquid; or as an oil-in-water liquid emulsion ora water-in-oil liquid emulsion. Tenofovir alafenamide hemifumarate mayalso be presented as a bolus, electuary, or paste.

Tenofovir alafenamide hemifumarate is preferably administered as part ofa pharmaceutical composition or formulation. Such pharmaceuticalcomposition or formulation comprises tenofovir alafenamide hemifumaratetogether with one or more pharmaceutically acceptablecarriers/excipients, and optionally other therapeutic ingredients. Theexcipient(s)/carrier(s) must be “acceptable” in the sense of beingcompatible with the other ingredients of the formulation and notdeleterious to the patient. Excipients include, but are not limited to,substances that can serve as a vehicle or medium for tenofoviralafenamide hemifumarate (e.g., a diluent carrier). They may be enclosedin hard or soft shell gelatin capsules, may be compressed into tablets,or may be incorporated directly with the food of the patient's diet.

Accordingly, the tablets, troches, pills, capsules, and the like mayalso contain, without limitation, the following: a binder(s), such ashydroxypropyl cellulose, povidone, or hydroxypropyl methylcellulose; afiller(s), such as microcrystalline cellulose, pregelatinized starch,starch, mannitol, or lactose monohydrate; a disintegrating agent(s),such as croscarmellose sodium, cross-linked povidone, or sodium starchglycolate; a lubricant(s), such as magnesium stearate, stearic acid, orother metallic stearates; a sweetening agent(s), such as sucrose,fructose, lactose, or aspartame; and/or a flavoring agent(s), such aspeppermint, oil of wintergreen, or a cherry flavoring. When the unitdosage form is a capsule, it may contain, in addition to materials ofthe above types, a liquid carrier, such as a vegetable oil or apolyethylene glycol. Various other materials may be present as coatingsor to otherwise modify the physical form of the solid unit dosage form.For instance, tablets, pills, or capsules may be coated with gelatin,polymers, wax, shellac, or sugar and the like. Of course, any materialused in preparing any unit dosage form typically will bepharmaceutically acceptable and substantially nontoxic in the amountsemployed. In addition, tenofovir alafenamide hemifumarate may beincorporated into sustained-release preparations and devices.

For infections of the eye or other external tissues, e.g., mouth andskin, the pharmaceutical compositions are preferably applied as atopical ointment or cream containing tenofovir alafenamide hemifumaratein an amount of, for example, 0.01 to 10% w/w (including activeingredient in a range between 0.1% and 5% in increments of 0.1% w/w suchas 0.6% w/w, 0.7% w/w, etc.), preferably 0.2 to 3% w/w and mostpreferably 0.5 to 2% w/w. When formulated in an ointment, the activeingredient may be employed with either a paraffinic or a water-miscibleointment base. Alternatively, the active ingredient may be formulated ina cream with an oil-in-water cream base.

Pharmaceutical compositions suitable for topical administration in themouth include lozenges comprising tenofovir alafenamide hemifumarate ina flavored basis, for example, sucrose and acacia or tragacanth;pastilles comprising the active ingredient in an inert basis such asgelatin and glycerin, or sucrose and acacia; and mouthwashes comprisingthe active ingredient in a suitable liquid carrier.

Formulations for rectal administration may be presented as a suppositorywith a suitable base comprising, for example, cocoa butter or asalicylate.

Pharmaceutical formulations suitable for parenteral administration aresterile and include aqueous and nonaqueous injection solutions that maycontain antioxidants, buffers, bacteriostats, and solutes that renderthe formulation isotonic with the blood of the intended recipient; andaqueous and nonaqueous sterile suspensions that may include suspendingagents and thickening agents. The formulations may be presented inunit-dose or multi-dose containers, for example, sealed ampoules andvials with elastomeric stoppers, and may be stored in a freeze-dried(lyophilized) condition requiring only the addition of the sterileliquid carrier (e.g., water for injections) immediately prior to use.Injection solutions and suspensions may be prepared from sterilepowders, granules, and tablets of the kind previously described.

In addition to the ingredients particularly mentioned above, thepharmaceutical compositions/formulations may include other ingredientsconventional in the art, having regard to the type of formulation inquestion.

In another embodiment, there is provided veterinary compositionscomprising tenofovir alafenamide hemifumarate together with a veterinarycarrier therefor. Veterinary carriers are materials useful for thepurpose of administering the composition to cats, dogs, horses, rabbits,and other animals, and may be solid, liquid, or gaseous materials thatare otherwise inert or acceptable in the veterinary art and arecompatible with the active ingredient. These veterinary compositions maybe administered orally, parenterally, or by any other desired route.

The tenofovir alafenamide hemifumarate can be used to provide controlledrelease pharmaceutical formulations containing a matrix or absorbentmaterial and an active ingredient of the invention, in which the releaseof the active ingredient can be controlled and regulated to allow lessfrequent dosing or to improve the pharmacokinetic or toxicity profile ofthe compound. Controlled release formulations adapted for oraladministration, in which discrete units comprising a compounds of theinvention, can be prepared according to conventional methods.

Useful dosages of tenofovir alafenamide hemifumarate can be determinedby comparing in vitro activities, and the in vivo activities in animalmodels. Methods for the extrapolation of effective amounts/dosages inmice and other animals to therapeutically effective amounts/dosages inhumans are known in the art.

The amount of tenofovir alafenamide hemifumarate required for use intreatment will vary with several factors, including but not limited tothe route of administration, the nature of the condition being treated,and the age and condition of the patient; ultimately, the amountadministered will be at the discretion of the attendant physician orclinician. The therapeutically effective amount/dose of tenofoviralafenamide hemifumarate depends, at least, on the nature of thecondition being treated, any toxicity or drug interaction issues,whether the compound is being used prophylactically (e.g., sometimesrequiring lower doses) or against an active disease or condition, themethod of delivery, and the pharmaceutical formulation, and will bedetermined by the clinician using conventional dose escalation studies.

In one embodiment, the oral dose of tenofovir alafenamide hemifumaratemay be in the range from about 0.0001 to about 100 mg/kg body weight perday, for example, from about 0.01 to about 10 mg/kg body weight per day,from about 0.01 to about 5 mg/kg body weight per day, from about 0.5 toabout 50 mg/kg body weight per day, from about 1 to about 30 mg/kg bodyweight per day, from about 1.5 to about 10 mg/kg body weight per day, orfrom about 0.05 to about 0.5 mg/kg body weight per day. As a nonlimitingexample, the daily candidate dose for an adult human of about 70 kg bodyweight will range from about 0.1 mg to about 1000 mg, or from about 1 mgto about 1000 mg, or from about 5 mg to about 500 mg, or from about 1 mgto about 150 mg, or from about 5 mg to about 150 mg, or from about 5 mgto about 100 mg, and may take the form of single or multiple doses.

The pharmaceutical compositions described herein may further include oneor more therapeutic agents in addition to tenofovir alafenamidehemifumarate. In one specific embodiment of the invention, theadditional therapeutic agent can be selected from the group consistingof HIV protease inhibiting compounds, HIV nonnucleoside inhibitors ofreverse transcriptase, HIV nucleoside inhibitors of reversetranscriptase, HIV nucleotide inhibitors of reverse transcriptase, HIVintegrase inhibitors, and CCR5 inhibitors.

Therapeutic methods include administering tenofovir alafenamidehemifumarate to a subject/patient in need of the same as a therapeuticor preventative treatment. Thus, tenofovir alafenamide hemifumarate maybe administered to a subject/patient having a medical disorder or to asubject who may acquire the disorder. One of ordinary skill willappreciate that such treatment is given in order to ameliorate, prevent,delay, cure, and/or reduce the severity of a symptom or set of symptomsof a disorder (including a recurring disorder). The treatment may alsobe given to prolong the survival of a subject, e.g., beyond the survivaltime expected in the absence of such treatment. The medical disordersthat may be treated with tenofovir alafenamide hemifumarate includethose discussed herein, including without limitation, HIV infection andHBV infection.

The following are nonlimiting, illustrative Examples.

EXAMPLE 1

Tenofovir alafenamide monofumarate solids (5.0 g) and9-[(R)-2-[[(R)-[[(S)-1-(isopropoxycarbonyl)ethyl]amino]phenoxyphosphinyl]methoxy]propyl]adenine(GS-7339) monofumarate solids (0.75 g) were charged into 35 g MTBE at22° C. and the mixture was stirred for 1 hour. A slurry was formed andwas dried in a rotary evaporator. 58 g acetonitrile (ACN) was chargedinto the solids and the mixture was heated to reflux to dissolve thesolids. The resulting solution was allowed to cool naturally whileagitated. A slurry was formed, and the slurry was further cooled byice-water-bath. The solids were isolated by filtration and washed with 5g ACN. The solids were dried in a vacuum oven at 40° C. overnight. 5.52g off-white solids were obtained. The solids were analyzed by XRPD andfound to contain tenofovir alafenamide monofumarate, GS-7339monofumarate, and tenofovir alafenamide hemifumarate.

EXAMPLE 2 Preparation of Tenofovir Alafenamide Hemifumarate ViaSelective Crystallization

9-[(R)-2-[[[[(S)-1-(isopropoxycarbonyl)ethyl]amino]phenoxyphosphinyl]methoxy]propyl]adenineas a slurry in ACN (9.7 kg slurry, 13.8 wt %, a diastereomeric mixtureof 1.0 kg (2.10 mol, 1 mol equiv) of9-[(R)-2-[[(S)-[[(S)-1-(isopropoxycarbonyl)ethyl]amino]phenoxyphosphinyl]methoxy]propyl]adenineand 0.35 kg of9-[(R)-2-[[(R)-[[(S)-1-(isopropoxycarbonyl)ethyl]amino]phenoxyphosphinyl]methoxy]propyl]adeninewas charged into a reactor and rinsed forward with dichloromethane (5kg). The mixture was concentrated under vacuum to about 3 L with jackettemperature below 40° C. The concentrate was then coevaporated with ACN(6 kg) under vacuum to about 3 L with jacket temperature below 40° C.The concentrate was diluted with ACN (8.5 kg) and warmed to 40-46° C.The warm mixture was filtered into a second reactor and the filtrate wascooled to 19-25° C.

To the above solution was charged fumaric acid (0.13 kg, 1.12 mol, 0.542mole equiv) followed by ACN (1 kg), and the mixture was heated to 67-73°C. The hot mixture was transferred into a reactor via a polishingfilter, and then adjusted to 54-60° C. Seed crystals (5 g) of thehemifumarate form of tenofovir alafenamide were charged (for example,the mixture can be seeded with tenofovir alafenamide hemifumarate formedin Example 1 or a subsequent production), and the resulting mixture wasagitated at 54-60° C. for about 30 minutes. The mixture was cooled overa minimum of 4 hours to 0-6° C., and then agitated at 0-6° C. for aminimum of 1 hour. The resulting slurry was filtered and rinsed withchilled (0-6° C.) ACN (2 kg). The product was dried under vacuum below45° C. until loss on drying (LOD) and organic volatile impurities (OVI)limits were met (LOD≦1.0%, dichloromethane content ≦0.19%, acetonitrilecontent ≦0.19%) to afford the final compound of the hemifumarate form oftenofovir alafenamide as a white to off-white powder (typical yield isabout 0.95 kg). ¹H NMR (400 MHz, d6 DMSO): δ 1.06 (d, J=5.6 Hz, 3H),1.12-1.16 (m, 9H), 3.77 (dd, J=10.4, 11.6 Hz, 1H), 3.84-3.90 (m, 2H),3.94 (m, 1H), 4.14 (dd, J=6.8, 14.8 Hz, 1H), 4.27 (m, 1H), 4.85 (heptet,J=6.0 Hz, 1H), 5.65 (t, J=11.2 Hz, 1H), 6.63 (s, 1H), 7.05 (d. J=7.6 Hz,2H), 7.13 (t, J=7.2 Hz, 1H), 7.24 (s, 2H), 7.29 (t, J=7.6 Hz, 2H), 8.13(t, J=13.6 Hz, 2H), ³¹P NMR (162 MHz, d6 DMSO): δ 23.3.

EXAMPLE 3 Preparation of Tenofovir Alafenamide Hemifumarate

To a jacketed reactor equipped with overhead agitator, was charged9-[(R)-2-[[(S)-[[(S)-1-(isopropoxycarbonyl)ethyl]amino]phenoxyphosphinyl]methoxy]propyl]adenine(10 g), fumaric acid (1.22 g), and ACN (100 mL). The mixture was heatedto 70-75° C. to dissolve the solids. Any undissolved particulates wereremoved by filtration through a cartridge filter. The filtered solutionwas cooled to 60-65° C., and seeded with 1% (by weight) of tenofoviralafenamide hemifumarate. The slurry was aged for 30 minutes and cooledto 0-5° C. over 2 hours. The temperature was maintained for 1-18 hours,and the resulting slurry was filtered and washed with 2 ml of cold ACN(0-5° C.). The solids were dried under vacuum at 50° C. to provide thehemifumarate form of tenofovir alafenamide, which was characterized asdescribed below.

Characterization of Tenofovir Alafenamide Hemifumarate from Example 3

Tenofovir alafenamide hemifumarate from Example 3 consists of9-[(R)-2-[[(S)-[[(S)-1-(isopropoxycarbonyl)ethyl]amino]phenoxyphosphinyl]methoxy]propyl]adenineand one-half an equivalent of fumaric acid. Tenofovir alafenamidehemifumarate is anhydrous, nonhygroscopic, and has a DSC onset endothermof about 131° C.

X-Ray Powder Diffraction

The XRPD pattern of tenofovir alafenamide hemifumarate was obtained inthe following experimental setting: 45 KV, 45 mA, Kα1=1.5406 Å, scanrange 2.-40°, step size 0.0084°, counting time: 8.25 s. The XRPD patternfor tenofovir alafenamide hemifumarate is shown in FIG. 1. Thecharacteristic peaks include: 6.9±0.2°, 8.6±0.2°, 10.0±0.2°, 11.0±0.2°,12.2±0.2°, 15.9±0.2°, 16.3±0.2°, 20.2±0.2°, and 20.8±0.2°.

Single-Crystal X-Ray Diffraction

The crystal size was 0.32×0.30×0.20 mm³. The sample was held at 123 Kand the data was collected using a radiation source with a wavelength of0.71073 Å in the theta range of 1.59 to 25.39°. Conditions of, and datacollected from the single-crystal X-ray diffraction are shown in Table1.

TABLE 1 Single-Crystal X-ray Diffraction Empirical formula C₂₃H₃₁N₆O₇PFormula weight 534.50 Temperature 123(2) K Crystal size 0.32 × 0.30 ×0.20 mm³ Theta range for data collection 1.59 to 25.39° Wavelength0.71073 Å Crystal system Tetragonal Space group P4(2)2(1)2 Unit celldimensions a = 18.1185(12) Å α = 90° b = 18.1185(12) Å β = 90° c =17.5747(11) Å γ = 90° Volume 5769.4(6) Å³ Z  8 Density (calculated)1.231 g/cm³DSC Analysis

The DSC analysis was conducted using 2.517 mg of tenofovir alafenamidehemifumarate. It was heated at 10° C./min over the range of 40-200° C.The onset endotherm was found to be about 131° C. (FIG. 2).

TGA Data

The TGA data were obtained using 4.161 mg of tenofovir alafenamidehemifumarate. It was heated at 10° C./min over the range of 25-200° C.The sample lost 0.3% weight before melting (FIG. 3). It was determinedto be an anhydrous form.

DVS Analysis

DVS analysis was conducted using 4.951 mg of tenofovir alafenamidehemifumarate. The material was kept at 25° C. in nitrogen at humiditiesranging from 10% to 90% relative humidity; each step was equilibratedfor 120 minutes. The sorption isotherm is shown at FIG. 4. The materialwas found to be nonhygroscopic, and to absorb 0.65% water at a relativehumidity of 90%.

Purging of Diastereomeric Impurity

In the prior syntheses of tenofovir alafenamide, one of the majorimpurities is typically the diastereomer9-[(R)-2-[[(R)-[[(S)-1-(isopropoxycarbonyl)ethyl]amino]phenoxyphosphinyl]methoxy]propyl]adenine.The hemifumarate form of tenofovir alafenamide from Example 3 has anexceptional capability to purge this diastereomeric impurity, ascompared with the capability of the monofumarate form (described in U.S.Pat. No. 7,390,791). The data in Table 2 (below) demonstrates thattenofovir alafenamide hemifumarate (Batch 2) purged the diastereomericimpurity to less than one-tenth of the starting concentration, whereasthe monofumarate form of tenofovir alafenamide (Batch 1) only slightlypurged the diastereomeric impurity.

TABLE 2 Purging Capability Comparison Diastereo- meric Fumaric Impurityacid Diastereo- in charge meric Starting (mole Product Impurity in BatchMaterial Solvent equivalent) obtained Product 1 9.3% ACN 0.9Monofumarate  7.6% form 2 10.0% ACN 0.5 Hemifumarate 0.65% formChemical Stability

Chemical stability of the hemifumarate form of tenofovir alafenamide wascompared with the monofumarate form. As shown in Table 3 (below), underidentical conditions, the hemifumarate form of tenofovir alafenamide waschemically more stable and exhibited better long-term storage stability,with significantly less degradation (% Total Deg. Products) than themonofumarate form. Conditions evaluated include temperature, relativehumidity (RH), and the open or closed state of the container cap.

TABLE 3 Chemical Stability Comparison Monofumarate form Hemifumarateform Time % TA* % Total % TA % Total Storage Points Area Deg. Area Deg.Condition (weeks) Normalized Products Normalized Products 40° C./ 0 97.10.69 98.4 0.05 75% RH 1 97.0 0.87 98.4 0.14 Cap 2 96.6 1.18 98.5 0.14Closed 4 96.4 1.49 98.4 0.25 8 95.4 2.36 98.0 0.49 40° C./ 0 97.1 0.6998.4 0.05 75% RH 1 96.9 0.90 98.5 0.15 Cap 2 96.6 1.10 98.5 0.14 Open 496.2 1.67 98.4 0.26 8 95.0 2.74 98.1 0.50 70° C. 0 97.1 0.69 98.4 0.05Cap 2 96.2 1.83 98.5 0.22 Closed 4 93.3 4.78 98.4 0.33 *TA is tenofoviralafenamideThermodynamic Stability

Stable form screening of tenofovir alafenamide hemifumarate showed thatit is thermodynamically stable in most solvents, such as ACN, toluene,ethyl acetate, methyl tert-butyl ether (MTBE), acetone, THF, and2-methyl THF. A similar stable form screening of the monofumarate formshowed that this form is not thermodynamically stable in theabove-listed solvents. When suspended in these solvents, themonofumarate form of tenofovir alafenamide fully converts to thehemifumarate form in THF and 2-methyl THF, and partially converts to thehemifumarate form in ACN, ethyl acetate, MTBE, and acetone, as well asat ambient temperatures.

Thermal Stability

As shown by the DSC data, the hemifumarate form of tenofovir alafenamidehas a melting point that is about 10° C. higher than that of themonofumarate form, indicating that the hemifumarate form has improvedthermal stability as compared with the monofumarate form.

All publications, patents, and patent documents are incorporated byreference herein, as though individually incorporated by reference. Theinvention has been described with reference to various specific andpreferred embodiments and techniques. However, it should be understoodthat many variations and modifications may be made while remainingwithin the spirit and scope of the invention.

What is claimed is:
 1. A composition comprising tenofovir alafenamidehemifumarate, wherein the composition comprises less than about 5% byweight of tenofovir alafenamide monofumarate.
 2. The composition ofclaim 1, wherein the composition comprises less than about 1% by weightof tenofovir alafenamide monofumarate.
 3. The composition of claim 1,wherein the composition comprises less than about 0.5% by weight oftenofovir alafenamide monofumarate.
 4. The composition of claim 1,wherein the ratio of fumaric acid to tenofovir alafenamide in saidcomposition is 0.5±0.1.
 5. The composition of claim 1, wherein the ratioof fumaric acid to tenofovir alafenamide in said composition is0.5±0.01.
 6. The composition of claim 1, having an X-ray powderdiffraction pattern that comprises 2theta values of 6.9±0.2° and8.6±0.2°.
 7. A pharmaceutical composition comprising the composition ofclaim 1 and a pharmaceutically acceptable excipient.
 8. Thepharmaceutical composition of claim 7, further comprising an additionaltherapeutic agent.
 9. The pharmaceutical composition of claim 8, whereinthe additional therapeutic agent is selected from the group consistingof human immunodeficiency virus (HIV) protease inhibiting compounds, HIVnonnucleoside inhibitors of reverse transcriptase, HIV nucleosideinhibitors of reverse transcriptase, HIV nucleotide inhibitors ofreverse transcriptase, HIV integrase inhibitors, and CCR5 inhibitors.10. A method for treating a human immunodeficiency virus (HIV) infectioncomprising administering to a subject in need thereof a therapeuticallyeffective amount of the composition of claim
 1. 11. A method fortreating an HIV infection comprising administering to a subject in needthereof a therapeutically effective amount of the pharmaceuticalcomposition of claim
 7. 12. The method for treating an HIV infection ofclaim 10, further comprising administering to the subject one or moreadditional therapeutic agents selected from the group consisting ofhuman immunodeficiency virus (HIV) protease inhibiting compounds, HIVnonnucleoside inhibitors of reverse transcriptase, HIV nucleosideinhibitors of reverse transcriptase, HIV nucleotide inhibitors ofreverse transcriptase, HIV integrase inhibitors, and CCR5 inhibitors.13. A method for treating a hepatitis B virus (HBV) infection comprisingadministering to a subject in need thereof a therapeutically effectiveamount of the composition of claim
 1. 14. A method for treating an HBVinfection comprising administering to a subject in need thereof atherapeutically effective amount of the pharmaceutical composition ofclaim
 7. 15. A method for preparing a pharmaceutical compositioncomprising combining the composition of claim 1 and a pharmaceuticallyacceptable excipient to provide the pharmaceutical composition.
 16. Themethod for treating an HIV infection of claim 10, wherein thecomposition is administered in multiple daily doses.
 17. The method fortreating an HIV infection of claim 10, wherein the composition isadministered in a single daily dose.
 18. The method for treating an HBVinfection of claim 13, wherein the composition is administered inmultiple daily doses.
 19. The method for treating an HBV infection ofclaim 13, wherein the composition is administered in a single dailydose.